生質燃料乃是指生物質經由轉換所獲得之能源(如生質乙醇、丁醇、生質柴油與生質氫氣)，至今仍備受重視，其中第三代生質料源—微藻亦是未來綠色替代能源的焦點。本研究希冀能建立進行微藻混營培養程序結合產物同步移除之整合型微藻丁醇醱酵系統，建構一套低污染(去除放流水中丁酸與乳酸)且高效能的永續循環之程序，以生產丁醇該生質燃料為標的。
為克服丁醇對Clostridium aectobutylicum ATCC824所產生的抑制作用，在發酵過程中亦使用同步移除。在以懸浮細胞進行批次發酵中，利用油醇為溶劑進行液液萃取，再結合氮氣氣提可顯著提升葡萄糖利用率及丁醇生產量。結果可得到葡萄糖利用率、丁醇生產速率、丁醇產率、選擇性分別為99 %、0.31 g/L/h、0.39 mol butanol/mol glucose及24.7。而採用固定化技術亦可提升C. acetobutylicum 之細胞濃度，本研究亦採固定化之C. acetobutylicum細胞進行產物同步移除發酵。以液液萃取結合氣提所得之結果在葡糖糖利用率99 %、丁醇生產速率0.545 g/L/h、產率0.34 mol butanol/mol glucose及選擇性15。
利用酸水解之微藻碳水化合物(主要為葡萄糖與木糖)之可行性進行丁醇發酵亦被評估，在以C. acetobutylicum懸浮細胞進行60 g/L藻糖之批式發酵中，發酵最終結果分別為丁醇濃度10.6 g/L丁醇生產速率1.04 g/L/h 及產率0.41 mol butanol/mol glucose。且以藻糖與一般葡萄糖為料源比較其發酵結果，利用藻糖可提升丁醇濃度12%，生產速率提升20%，產率亦提升17%。
而以73.5 g/L濃縮藻糖為料源，進行固定化細胞丁醇發酵結合產物同步移除，可得發酵最終結果分別為丁醇濃度12.01 g/L丁醇生產速率0.44 g/L/h 及產率0.41 mol butanol/mol glucose及選擇性6.66。本實驗亦可作為在60 g/L藻糖濃度下，連續式丁醇發酵結合產物同步移除之前測。
由於丁醇發酵最終產物內含有揮發性有機酸，為降低發酵液COD及達成廢水循環再利用，挑選藻株C. vulgaris JSC6進行混營培養之測試，得到最佳培養條件為5%發酵液混合Basal培養基，進而生產微藻生物質濃度達6.81 g/L，藻體所累積之29%碳水化合物將可作為後續醱酵製程之料源。以微藻生質體(水解藻糖)為料源結合液液萃取與氣提之產物同步移除整合型微藻丁醇醱酵系統，可建構一套低汙染且高效能的零碳排且永續循環之程序。

Biofuels such as alcohols, biodiesel, and biohydrogen derived from biomass have received much attention due to their potential to serve as alternative renewable energy resources for energy security and sustainability. In particular, using third generation microalgal biomass as feedstock for biofuels production has recently been regarded with great interest. In this study, we focused on developing a low-pollution, sustainable and highly efficient microalgae-based bio-butanol production system with simultaneous removal of organic acids from the butanol fermentation effluent.
In situ product removal has been used to overcome the product inhibition effect of butanol on Clostridium aectobutylicum ATCC824 in batch butanol fermentation. A combination of liquid-liquid extraction using oleyl alcohol and gas stripping with nitrogen gas greatly improved the glucose utilization and butanol production in batch butanol fermentation using suspended cells. In free cell batch fermentation, the glucose utilization, butanol productivity, butanol yield and selectivity were improved to 99 %, 0.31 g/L/h, 0.39 mol butanol/mol glucose and 24.7, respectively. Immobilization of C. acetobutylicum was done to enhance the cell concentration in the reactor and immobilized cells were used in butanol fermentation integrated with in situ product removal techniques (gas stripping, liquid liquid extraction + gas stripping). Two immobilized cell batch fermentation tests were conducted, and the results of liquid-liquid extraction combined with gas stripping show that glucose utilization, butanol productivity, butanol yield and selectivity were 99 %, 0.545 g/L/h, 0.34 mol butanol/mol glucose and 15.
The feasibility of using microalgal carbohydrates as obtained by simple acid hydrolysis (primarily glucose and xylose) was evaluated in batch fermentation using suspended cells of C. acetobutylicum. The maximum butanol concentration, maximum productivity and yield with 60 g/L microalgal sugars were 10.6 g/L, 1.04 g/L/h and 0.41 mol butanol/mol glucose, respectively. The fermentation performance using microalgal sugar is better than that using commercial glucose. The maximum butanol concentration, maximum productivity and yield were improved by 12, 20, and 17%, respectively. The integrated system combining in situ product removal (liquid-liquid extraction/gas stripping) with immobilized cells using hydrolyzed microalgal biomass was conducted in batch mode. The maximum butanol concentration, maximum productivity, yield and selectivity using 73.5 g/L concentrated microalgal sugars were 12.01 g/L, 0.44 g/L/h and 0.41 mol butanol/mol glucose and 6.66. The same experimental setup can be adapted for CSTR mode of fermentation with 60 g/L microalgal sugar and in situ product removal.
Finally, to reduce the COD of the fermentation waste and to reuse the fermentation broth, Chlorella vulgaris JSC-6 was cultivated in butanol fermentation broth in mixotrophic mode of cultivation. We found that 5% fermentation broth mixed with Basal medium yielded the best biomass production (6.81 g/L). The obtained biomass containing 29% carbohydrates can further be hydrolyzed and reused as the feedstock for biobutanol fermentation.
A combination of liquid-liquid extraction and gas stripping integrated with butanol fermentation system using microalgal sugars can help achieve a low-pollution and highly-efficient microalgae-based bio-butanol production system, which is not only free of carbon dioxide emissions but is also sustainable.

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